BackgroundThe vapor phase of the volatile pyrethroid transfluthrin incapacitates mosquitoes and prevents them from feeding. Although existing emanator products for delivering volatile pyrethroids protect against outdoor mosquito bites, they are too short-lived to be practical or affordable for routine use in low-income settings. New transfluthrin emanators, comprised simply of treated hessian fabric strips, have recently proven highly protective against outdoor-biting vectors of lymphatic filariasis, arboviruses and malaria, but their full protective lifespan, minimum dose requirements, and range of protection have not previously been assessed.MethodologyThe effects of transfluthrin-treated hessian strips upon mosquito biting exposure of users and nearby non-users, as well as dependence of protection upon treatment dose, were measured outdoors in rural Tanzania using human landing catches (HLC).Principal findingsStrips treated with 10ml of transfluthrin prevented at least three quarters (p < 0.001) of outdoor bites by Anopheles arabiensis, Culex spp. and Mansonia spp. mosquitoes, and >90% protection against bites on warmer nights with higher evaporation rates, for at least one year. Strips treated with this high dose also reduced biting exposure of non-users at a distance of up to 5m from the strips for An. arabiensis (p < 0.001) and up to 2m for Mansonia spp. (p = 0.008), but provided no protection to non-users against Culex spp. No evidence of increased risk for non-users, caused by diversion of mosquitoes to unprotected individuals, was found at any distance within an 80m radius. A dose of only 1ml provided equivalent protection to the 10ml dose against An. arabiensis, Culex spp. and Mansonia spp. mosquitoes over 6 months (p < 0.001).Conclusions/SignificanceTransfluthrin-treated hessian emanators provide safe, affordable, long-term protection against several different pathogen-transmitting mosquito taxa that attack humans outdoors, where they are usually active and cannot be protected by bed nets or residual sprays with conventional, solid-phase insecticides.

We investigate the role of plastoquinone (PQ) diffusion in the control of the photosynthetic electron transport. A control analysis reveals an unexpected flux control of the whole chain electron transport by photosystem (PS) II. The contribution of PSII to the flux control of whole chain electron transport was high in stacked thylakoids (control coefficient, CJ(PSII) =0.85), but decreased after destacking (CJ(PSII)=0.25). From an 'electron storage' experiment, we conclude that in stacked thylakoids only about 50 to 60% of photoreducable PQ is involved in the light-saturated linear electron transport. No redox equilibration throughout the membrane between fixed redox groups at PSII and cytochrome (cyt) bf complexes, and the diffusable carrier PQ is achieved. The data support the PQ diffusion microdomain concept by Lavergne et al. [J. Lavergne, J.-P. Bouchaud, P. Joliot, Biochim. Biophys. Acta 1101 (1992) 13-22], but we come to different conclusions about size, structure and size distribution of domains. From an analysis of cyt b6 reduction, as a function of PSII inhibition, we conclude that in stacked thylakoids about 70% of PSII is located in small domains, where only 1 to 2 PSII share a local pool of a few PQ molecules. Thirty percent of PSII is located in larger domains. No small domains were found in destacked thylakoids. We present a structural model assuming a hierarchy of specific, strong and weak interactions between PSII core, light harvesting complexes (LHC) II and cyt bf. Peripheral LHCII's may serve to connect PSII-LHCII supercomplexes to a flexible protein network, by which small closed lipid diffusion compartments are formed. Within each domain, PQ moves rapidly and shuttles electrons between PSII and cyt bf complexes in the close vicinity. At the same time, long range diffusion is slow. We conclude, that in high light, cyt bfcomplexes located in distant stromal lamellae (20 to 30%) are not involved in the linear electron transport.

SummaryPlasmodium berghei is the causative agent of rodent malaria and is widely used as a model system to study the liver stage of Plasmodium parasites. The entry of P. berghei sporozoites into hepatocytes has extensively been studied, but little is known about parasite-host interaction during later developmental stages of the intracellular parasite. Growth of the parasite far beyond the normal size of the host cell is an important stress factor for the infected cell. Cell stress is known to trigger programmed cell death (apoptosis) and we examined several apoptotic markers in P. berghei -infected cells and compared their level of expression and their distribution to that of non-infected cells. As none of the apoptotic markers investigated were found altered in infected cells, we hypothesized that parasite infection might confer resistance to apoptosis of the host cell. Treatment with peroxide or serum deprivation induced apoptosis in non-infected HepG2 cells, whereas P. bergheiinfected cells appeared protected, indicating that the parasite interferes indeed with the apoptotic machinery of the host cell. To prove the physiological relevance of these results, mice were infected with high numbers of P. berghei sporozoites and treated with tumour necrosis factor (TNF)-α α α α / D -galactosamine to induce massive liver apoptosis. Liver sections of these mice, stained for degraded DNA, confirmed that infected cells containing viable parasites were protected from programmed cell death. However, in nontreated control mice as well as in TNF-α α α α -treated mice a small proportion of dead intracellular parasites with degraded DNA were detected. Most hepatocytes containing dead parasites provoked an infiltration of immunocompetent cells, indicating that these cells are no longer protected from cell death.

SummaryCysteine proteases mediate liberation of Plasmodium berghei merozoites from infected hepatocytes. In an attempt to identify the responsible parasite proteases, we screened the genome of P. berghei for cysteine protease-encoding genes. RT-PCR analyses revealed that transcription of four out of five P. berghei serine repeat antigen (PbSERA) genes was strongly upregulated in late liver stages briefly before the parasitophorous vacuole membrane ruptured to release merozoites into the host cell cytoplasm, suggesting a role of PbSERA proteases in these processes. In order to characterize PbSERA3 processing, we raised an antiserum against a nonconserved region of the protein and generated a transgenic P. berghei strain expressing a TAP-tagged PbSERA3 under the control of the endogenous promoter. Immunofluorescence assays revealed that PbSERA3 leaks into the host cell cytoplasm during merozoite development, where it might contribute to host cell death or activate host cell proteases that execute cell death. Importantly, processed PbSERA3 has been detected by Western blot analysis in cell extracts of schizont-infected cells and merozoiteinfected detached hepatic cells.

Evidence obtained during recent years suggests that BMyb, a highly conserved member of the Myb transcription factor family, plays a key role in cell proliferation. We have shown previously that the activity of B-Myb is stimulated by cyclin A/Cdk2-dependent phosphorylation of the carboxyl-terminus of B-Myb. We have now investigated in more detail the eect of other cyclins on B-Myb. Here, we show that cyclin D1, in contrast to cyclin A, strongly inhibits the activity of B-Myb. This inhibitory eect does not involve increased phosphorylation of B-Myb but seems to rely on the formation of a speci®c complex of B-Myb and cyclin D1. Our work identi®es B-Myb as an interacting partner for cyclin D1 and suggest that the activity of B-Myb during the cell cycle is controlled by the antagonistic eects of cyclin D1 and A. The results presented here suggest a more general role of cyclin D1 as regulator of transcription in addition to the known eect on RB phosphorylation.